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University of Stuttgart

10 ARTICLES PUBLISHED IN JoVE

Biology

In Vitro Analysis of PDZ-dependent CFTR Macromolecular Signaling Complexes

Yanning Wu¹, Shuo Wang¹, Chunying Li^1,2,3

¹Department of Biochemistry & Molecular Biology, Wayne State University School of Medicine, ²Cardiovascular Research Institute, Wayne State University School of Medicine, ³Barbara Ann Karmanos Cancer Institute, Wayne State University School of Medicine

Cystic fibrosis transmembrane conductance regulator (CFTR), an epithelial chloride channel, has been reported to interact with various proteins and regulate important cellular processes; among them the CFTR PDZ motif-mediated interactions have been well documented. This protocol describes methods we developed to assemble a PDZ-dependent CFTR macromolecular signaling complex in vitro.

Behavior

Creating Dynamic Images of Short-lived Dopamine Fluctuations with lp-ntPET: Dopamine Movies of Cigarette Smoking

Evan D. Morris^1,2,3,4, Su Jin Kim^1,3, Jenna M. Sullivan^1,3,4, Shuo Wang^3,4, Marc D. Normandin⁵, Cristian C. Constantinescu⁶, Kelly P. Cosgrove^1,2,3

¹Diagnostic Radiology, Yale University, ²Psychiatry, Yale University, ³Yale PET Center, Yale University, ⁴Biomedical Engineering, Yale University, ⁵Nuclear Medicine, Massachusetts General Hospital, ⁶Radiological Sciences, University of California, Irvine

We present a novel PET imaging approach for capturing dopamine fluctuations induced by cigarette smoking. Subjects smoke in the PET scanner. Dynamic PET images are modeled voxel-by-voxel in time by lp-ntPET, which includes a time-varying dopamine term. The results are 'movies' of dopamine fluctuations in the striatum during smoking.

Engineering

How to Ignite an Atmospheric Pressure Microwave Plasma Torch without Any Additional Igniters

Martina Leins¹, Sandra Gaiser¹, Andreas Schulz¹, Matthias Walker¹, Uwe Schumacher¹, Thomas Hirth¹

¹Institute of Interfacial Process Engineering and Plasma Technology, University of Stuttgart

This movie shows how an atmospheric plasma torch can be ignited by microwaves with no additional igniters and provides a stable and continuous plasma operation suitable for plenty of applications.

Environment

Optimized Procedure for Determining the Adsorption of Phosphonates onto Granular Ferric Hydroxide using a Miniaturized Phosphorus Determination Method

Eduard Rott¹, Tobias Reinhardt¹, Stephan Wasielewski¹, Ellen Raith-Bausch¹, Ralf Minke¹

¹Institute for Sanitary Engineering, Water Quality and Solid Waste Management, University of Stuttgart

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric hydroxide, with little effort and high reliability. In a buffered solution, the phosphonate is brought into contact with the adsorbent using a rotator and then analyzed via a miniaturized phosphorus determination method.

JoVE Journal

A 100 KW Class Applied-field Magnetoplasmadynamic Thruster

Baojun Wang¹, Haibin Tang², Yibai Wang¹, Chao Lu¹, Cheng Zhou³, Yangyang Dong¹, Ge Wang³, Yuntian Cong³, Daniel Luu⁴, Jinbin Cao²

¹Key Laboratory of Spacecraft Design Optimization & Dynamic Simulation Technologies of Ministry of Education, School of Astronautics, Beihang University, ²Key Laboratory of Spacecraft Design Optimization & Dynamic Simulation Technologies of Ministry of Education, School of Space and Environment, Beihang University, ³Beijng Institute of Control Engineering, ⁴School of Aerospace Engineering and Geodesy, University of Stuttgart

The goal of this protocol is to introduce the design of a 100 kW class applied-field magnetoplasmadynamic thruster and relevant experimental methods.

Chemistry

Synthesis of Soft Polysiloxane-urea Elastomers for Intraocular Lens Application

Natascha Riehle^*1,2, Sibylle Thude^*3, Andreas Kandelbauer^1,2, Günter E. M. Tovar^3,4, Günter Lorenz^1,2

¹Reutlingen Research Institute, Reutlingen University, ²School of Applied Chemistry, Reutlingen University, ³Fraunhofer-Institute for Interfacial Engineering and Biotechnology IGB, ⁴Institute of Interfacial Process Engineering and Plasma Technology IGVP, University of Stuttgart

This study describes synthetic routes for aminopropyl-terminated polydimethylsiloxanes and polydimethyl-methyl-phenyl-siloxane-block copolymers and for soft polysiloxane-based urea (PSU) elastomers. It presents the application of PSUs as accommodating an intraocular lens. An evaluation method for in vitro cytotoxicity is also described.

Behavior

Simultaneous Eye Tracking and Single-Neuron Recordings in Human Epilepsy Patients

Shuo Wang¹, Nand Chandravadia², Adam N. Mamelak², Ueli Rutishauser^2,3,4

¹Department of Chemical and Biomedical Engineering, and Rockefeller Neuroscience Institute, West Virginia University, ²Departments of Neurosurgery and Neurology, Cedars-Sinai Medical Center, ³Center for Neural Science and Medicine, Department of Biomedical Sciences, Cedars-Sinai Medical Center, ⁴Division of Biology and Biological Engineering, California Institute of Technology

We describe a method to conduct single-neuron recordings with simultaneous eye tracking in humans. We demonstrate the utility of this method and illustrate how we used this approach to obtain neurons in the human medial temporal lobe that encode targets of a visual search.

Cancer Research

Detection of Lung Tumor Progression in Mice by Ultrasound Imaging

Nour Ghaddar^1,2, Shuo Wang¹, Véronique Michaud¹, Urszula Kazimierczak^1,3, Nicolas Ah-son¹, Antonis E. Koromilas^1,4

¹Lady Davis Institute for Medical Research, Sir Mortimer B. Davis-Jewish General Hospital, ²Division of Experimental Medicine, Faculty of Medicine, McGill University, ³Department of Cancer Immunology, Chair of Medical Biotechnology, Poznan University of Medical Sciences, ⁴Gerald Bronfman Department of Oncology, Faculty of Medicine, McGill University

This protocol describes the steps taken to induce KRAS lung tumors in mice as well as the quantification of formed tumors by ultrasound imaging. Small tumors are visualized in early timepoints as B-lines. At later timepoints, relative tumor volume measurements are achieved by the measurement tool in the ultrasound software.

Medicine

Hemocompatibility Testing of Blood-Contacting Implants in a Flow Loop Model Mimicking Human Blood Flow

Antonia Link¹, Giorgio Cattaneo^2,3, Eduard Brynda⁴, Tomas Riedel⁴, Johanka Kucerova⁴, Christian Schlensak¹, Hans Peter Wendel¹, Stefanie Krajewski¹, Tatjana Michel¹

¹Department of Thoracic, Cardiac and Vascular Surgery, University Hospital Tuebingen, ²Acandis GmbH, ³Institute of Biomedical Engineering, University of Stuttgart, ⁴Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic

This protocol describes a comprehensive hemocompatibility evaluation of blood-contacting devices using laser-cut neurovascular implants. A flow loop model with fresh, heparinized human blood is applied to mimic blood flow. After perfusion, various hematologic markers are analyzed and compared to the values gained directly after blood collection for hemocompatibility evaluation of the tested devices.

Biochemistry

Single-Molecule Imaging of Lateral Mobility and Ion Channel Activity in Lipid Bilayers using Total Internal Reflection Fluorescence (TIRF) Microscopy

Shuo Wang¹, Stephan Nussberger¹

¹Biophysics Department, Institute of Biomaterials and Biomolecular Systems, University of Stuttgart

This protocol describes how to use TIRF microscopy to track individual ion channels and determine their activity in supported lipid membranes, thereby defining the interplay between lateral membrane movement and channel function. It describes how to prepare the membranes, record the data, and analyze the results.